Case Report: Sustained Efficacy of Lumasiran at 18 Months in Primary Hyperoxaluria Type 1

Background: Primary hyperoxaluria type 1 (PH1) is a rare genetic disease caused by hepatic overproduction of oxalate, ultimately responsible for kidney stones, kidney failure and systemic oxalosis. Lumasiran, is a liver-directed RNA interference therapeutic agent. It has been shown to reduce hepatic oxalate production by targeting glycolate oxidase, and to dramatically reduce oxalate excretion. Care Report: We present the case of a teenager patient affected by PH1, who entered in the lumasiran compassionate use program. The patient had a rapid and sustained decrease in urinary oxalate/creatinine ratio, with a mean reduction after lumasiran administration of about 70%. During the 18 months long follow-up, urinary oxalate remained low, reaching nearly normal values. Plasma oxalate also decreased dramatically. Normal levels were reached immediately after the first dose and remained consistently low thereafter. During the same follow-up period, eGFR remained stable at about 60 ml/min/1.73 m2, but no new kidney stones were observed. Existing kidney stones did not increase in size. The patient did not suffer renal colic events and did not require further urological interventions. Conclusion: In our severely affected PH1 patient, lumasiran proved to be very effective in rapidly and consistently reducing plasma oxalate and urinary excretion to normal and near-normal levels, respectively. In the 18 months long follow-up post-lumasiran, the eGFR remained stable and the patient showed clinical improvements. As far as we know, this report covers the longest observation period after initiation of this novel RNAi therapy.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Accuracy of automated flow cytometry-based leukocyte counts to rule out urinary tract infection in febrile children: A prospective cross-sectional study

Automated flow cytometry of urine remains an incompletely validated method to rule out urinary tract infection (UTI) in children. This cross-sectional analytical study was performed to compare the predictive values of flow cytometry and a dipstick test as initial diagnostic tests for UTI in febrile children and prospectively included 1,106 children (1,247 episodes). Urine culture was used as the gold standard test for diagnosing UTI. The performance of screening tests to diagnose UTI were established using receiver operating characteristic (ROC) analysis. Among these 1,247 febrile episodes, 221 UTIs were diagnosed (17.7% [95% confidence interval {CI}, 15.6 to 19.8%]). The area under the ROC curve for flow cytometry white blood cell (WBC) counts (0.99 [95% CI, 0.98 to 0.99]) was significantly superior to that for red blood cell (0.74 [95% CI, 0.70 to 0.78]) and bacterial counts (0.89 [95% CI, 0.87 to 0.92]) (P<0.001). Urinary WBC counts also had a significantly higher area under the ROC curve than that of the leukocyte esterase (LE) dipstick (0.92 [95% CI, 0.90 to 0.94]), nitrite dipstick (0.83 [95% CI, 0.80 to 0.87]), or the combination of positive LE and/or nitrite dipstick (0.91 [95% CI, 0.89 to 0.93]) test (P<0.001). The presence of ≥35 WBC/μl of urine was the best cutoff point, yielding both a high sensitivity (99.5% [95% CI, 99 to 100%]) and an acceptable specificity (80.6% [95% CI, 78 to 83%]). Using this cutoff point would have reduced the number of samples sent to the laboratory for culture by 67%. In conclusion, the determination of urinary WBC counts by flow cytometry provides optimal performance as an initial diagnostic test for UTI in febrile children.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Rhabdomyolysis and Acute Kidney Injury as Leading COVID-19 Presentation in an Adolescent.

Severe acute respiratory syndrome coronavirus 2, the virus responsible of the current COVID-19 pandemic, has limited impact in the pediatric population. Children are often asymptomatic or present mild flu-like symptoms. We report the case of a COVID-19-infected adolescent presenting severe rhabdomyolysis and acute kidney injury without any fever or respiratory symptoms.info:eu-repo/semantics/publishe

Gender Differences in Hepatology Medical Literature

Studies suggest that gender differences in academic medicine exist. Men frequently have better measures of performance such as number of publications, number of citations, remuneration, and funding. To evaluate whether a gender disparity in authorship exists. We recorded the gender of first and senior authors of original papers, editorials/reviews from liver-related manuscripts in Gastroenterology, Hepatology, Transplantation, American Journal of Gastroenterology, and Liver Transplantation from January 2014 to 2016. Of 2424 articles reviewed, we excluded 232 (10%) due to inability to determine gender. Among papers analyzed, 72.0% were original and 28.1% reviews/editorials with 65.1% of first authors being male and 34.9% female. Only 20.3% of papers with multiple authors had a female senior author. The proportion of male first and senior authorship between original papers and reviews/editorials was comparable. 72% of original papers had a male as first or senior author, but only 28% females. 71% of review/editorial papers had a male as first or senior author, but only 29% females. When the senior author of an original paper was female, 47.1% of first authors were male and 52.9% female. When the senior author was male, 67.1% of first authors were male and 32.9% female (p < 0.00001). A significant gender difference exists in Hepatology publications. Female authorship mirrors the percentage of female AASLD membership; however, female senior authorship remains disproportionate. In general, funding for male authors is greater. Fewer women are first authors when the senior author is male, highlighting the importance of female mentorship in Hepatology

BK Polyomavirus in Pediatric Renal Transplantation—What We Know and What We Do Not

BK polyomavirus (BKPyV) is still a real threat in the management of kidney transplantation. Immunosuppressive treatment disrupts the equilibrium between virus replication and immune response, and uncontrolled BKPyV replication leads to nephropathy (BKPyV nephropathy). The first evidence of BKPyV reactivation in transplant recipients is the detection of viral shedding in urine, which appears in 20% to 60% of patients, followed by BKPyV viremia in 10–20% of kidney transplant recipients. BKPyV nephropathy eventually occurs in 1–10% of this population, mainly within the first 2 years post-transplantation, causing graft loss in about half of those patients. Few data exist regarding the pediatric population and we focus on them. In this paper, we review the existing diagnostic methods and summarize the evidence on the role of BKPyV humoral and cellular immunity in modulating the clinical course of BKPyV infection and as potential predictors of the outcome. We look at the known risk factors for BKPyV nephropathy in the immunosuppressed patient. Finally, we propose a sensible clinical attitude in order to screen and manage BKPyV infection in kidney transplant children

Genome-wide RNAi screen reveals a new role of a WNT/CTNNB1 signaling pathway as negative regulator of virus-induced innate immune responses.

To identify new regulators of antiviral innate immunity, we completed the first genome-wide gene silencing screen assessing the transcriptional response at the interferon-β (IFNB1) promoter following Sendai virus (SeV) infection. We now report a novel link between WNT signaling pathway and the modulation of retinoic acid-inducible gene I (RIG-I)-like receptor (RLR)-dependent innate immune responses. Here we show that secretion of WNT2B and WNT9B and stabilization of β-catenin (CTNNB1) upon virus infection negatively regulate expression of representative inducible genes IFNB1, IFIT1 and TNF in a CTNNB1-dependent effector mechanism. The antiviral response is drastically reduced by glycogen synthase kinase 3 (GSK3) inhibitors but restored in CTNNB1 knockdown cells. The findings confirm a novel regulation of antiviral innate immunity by a canonical-like WNT/CTNNB1 signaling pathway. The study identifies novel avenues for broad-spectrum antiviral targets and preventing immune-mediated diseases upon viral infection

Pharmacological inhibition of GSK3 stabilizes an active CTNNB1 pool and negatively regulates antiviral innate immunity.

<p>(A–B) Relative fold induction of <i>IFNB1</i>-driven reporter activity (A) and fold induction of CTNNB1-TCF/LEF dependent reporter activity TOP-flash (B) in HEK 293T cells transduced with lentivirus-expressing shRNA NT (control) or shRNA 45 targeting <i>CTNNB1</i> for four days and subjected to GSK3 inhibition with BIO (5 µM) or BIO-acetoxime (10 µM) and SeV infection for 16 hours. (C) Immunoblot analysis of autophosphorylated GSK3α/β at Tyr279/216, dephosphorylated active CTNNB1 at Ser37/Thr41 and IFIT1 in HEK 293T cells treated with GSK3 inhibitors BIO (5 µM) or BIO-acetoxime (10 µM) and infected with SeV for 16 hours. (D) Immunoblot analysis of dephosphorylated active CTNNB1 at Ser37/Thr41, IFIT1 and phosphorylated NFKBIA (IκBα) at Ser32 in SeV-infected HEK 293T cells treated as described in (A–B) and treated with BIO (5 µM) or BIO-acetoxime (10 µM). (E) Relative fold induction of TOP-flash, <i>IFNB1</i>, <i>ISG56</i>, <i>NF-κB</i> and <i>EF1α</i>-driven reporter activity in HEK 293T cells subjected to GSK3 inhibition with BIO (5 µM), TCF/LEF interaction inhibition with PNU74654 (6 µM) or both inhibitors prior to SeV infection for 16 hours.</p

Cellular map of prioritized gene hits as potential modulators of innate immunity.

<p>Cellular mapping of gene products of 96 gene hits out of 114 that modulated endogenous <i>IFNB1</i> expression either as Positive Regulators (71 PRs: Red) or as Negative Regulators (25 NRs: green) of the signaling pathway following SeV infection. PRs and NRs were positioned on a cell map and classified according to sub-cellular compartments and cellular function using annotation information from Gene Ontology, KEGG pathway, Uniprot, NCBI and Ingenuity pathway Analysis (IPA). The four functional groups are represented by different symbols: I - SeV specific (triangle), II - cytoplasmic dsRNA sensing (diamond), III - MAVS-dependent signaling (square) and IV - Nuclear import or transcription factor-dependent process (circle). Gene hits with knockdown efficiency confirmed by qRT-PCR with at least two independent shRNAs are highlighted in bold (41 PRs and 9NRs). The RLR and WNT signaling pathways are represented in light and dark blue respectively. Virus-induced activation of the WNT/CTNNB1 canonical-like pathway in feedback inhibition of antiviral innate responses is indicated in red.</p

CTNNB1 is a negative regulator of antiviral innate immunity.

<p>(A) <i>IFNB1</i> promoter-driven luciferase activity in HEK 293T cells transduced with lentivirus-expressing shRNA NT (control) or 3 shRNAs targeting CTNNB1 for four days and subjected to SeV infection for 16 hours. (B) Immunoblot analysis of CTNNB1 and IFIT1 in HEK 293T cells transduced with 3 lentivirus-expressing shRNAs targeting CTNNB1 and infected with SeV. (C) Immunoblot analysis of CTNNB1, IFIT1 and DDX58 in SeV-infected HEK 293T cells previously transduced with a shRNA or transfected with a pool of four siRNAs targeting CTNNB1 for three days. (D–F) Fold induction of <i>IFNB1</i> (D), <i>DDX58</i> (E) and <i>TNF</i> (F) mRNA levels in HEK 293T cells treated as described in (C). qRT-PCR determination represents the average mRNA RQ normalized versus ACTIN and HPRT1 mRNA. (G) ELISA quantification of secreted IFN-β protein in supernatant of HEK 293T cells treated as described in (A). (H–I) Fold induction of <i>IFNB1</i> promoter-driven luciferase activity (H) and of CTNNB1-TCF/LEF dependent reporter activity (TOP-flash, I) following dose-dependent transfection of CTNNB1-expressing plasmid (50, 100, 150 and 200 ng) for 48 hours in SeV-infected HEK 293T cells.</p